College of Chemistry and Molecular Sciences, Key Laboratory of Biomedical Polymers of Ministry of Education, Institute of Molecular Medicine, Renmin Hospital of Wuhan University, School of Microelectronics, Wuhan University, Wuhan 430072, P. R. China.
Molecular Science and Biomedicine Laboratory (MBL), State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, P. R. China.
ACS Sens. 2023 May 26;8(5):2096-2104. doi: 10.1021/acssensors.3c00453. Epub 2023 May 17.
The large-scale pandemic and fast evolution of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants have triggered an urgent need for an efficient and sensitive on-site nucleic acid testing method with single-nucleotide polymorphism (SNP) identification capability. Here, we report a multiplexed electrical detection assay based on a paperclip-shaped nucleic acid probe (PNprobe) functionalized field-effect transistor (FET) biosensor for highly sensitive and specific detection and discrimination of SARS-CoV-2 variants. The three-stem structure of the PNprobe significantly amplifies the thermodynamic stability difference between variant RNAs that differ in a single-nucleotide mutation. With the assistance of combinatorial FET detection channels, the assay realizes simultaneously the detection and identification of key mutations of seven SARS-CoV-2 variants, including nucleotide substitutions and deletions at single-nucleotide resolution within 15 min. For 70 simulated throat swab samples, the multiplexed electrical detection assay shows an identification accuracy of 97.1% for the discrimination of SARS-CoV-2 variants. Our designed multiplexed electrical detection assay with SNP identification capability provides an efficient tool to achieve scalable pandemic screening.
大规模的大流行和严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2) 变体的快速演变，引发了对具有单核苷酸多态性 (SNP) 识别能力的高效和敏感的现场核酸检测方法的迫切需求。在这里，我们报告了一种基于回形针状核酸探针 (PNprobe) 功能化场效应晶体管 (FET) 生物传感器的多重电化学生物检测方法，用于高灵敏度和特异性检测和区分 SARS-CoV-2 变体。PNprobe 的三链结构显着放大了在单个核苷酸突变中存在差异的变体 RNA 的热力学稳定性差异。借助组合 FET 检测通道，该检测方法在 15 分钟内实现了对七种 SARS-CoV-2 变体的关键突变的同时检测和鉴定，包括单核苷酸分辨率下的核苷酸取代和缺失。对于 70 个模拟的咽喉拭子样本，多重电化学生物检测方法在区分 SARS-CoV-2 变体方面的识别准确率为 97.1%。我们设计的具有 SNP 识别能力的多重电化学生物检测方法提供了一种有效的工具，可实现可扩展的大流行筛查。
